LC-MS/MS检测生物检材中雷公藤甲素和雷公藤酯甲

Determination of Triptolide and Wilforlide A in Biological Samples by LC-MS/MS

目的建立生物检材中雷公藤甲素和雷公藤酯甲的液相色谱-串联质谱（LC-MS/MS）分析方法,并进行方法学验证。方法 0.4 m L血液、尿液或0.4 g肝组织加入内标混匀后用乙酸乙酯进行提取,提取物经Allure PFP Propyl柱（100 mm×2.1 mm,5μm）分离,以甲醇-20 mmol/L乙酸铵溶液梯度洗脱,采用电喷雾正离子化（ESI＋）、多反应监测检测雷公藤甲素和雷公藤酯甲。结果各生物检材中雷公藤甲素和雷公藤酯甲在相应的线性范围内线性良好（r〉0.995 0）,检出限均为2 ng/m L或2 ng/g,回收率为61.08%～102.98%,日内精密度和日间精密度均小于12.58%,准确度为90.61%～105.80%。结论所建方法简便、选择性好,适用于同时分析各种生物检材中的雷公藤甲素和雷公藤酯甲,为雷公藤中毒的法医学鉴定和临床诊治提供技术保障。

Objective To determinate triptolide and wilforlide A in biological samples by liquid chromatography-tandem mass spectrometry（LC-MS/MS） method and to verify the method.Methods After 0.4 m L blood,urine or 0.4 g hepatic tissues with internal standard were extracted by ethyl acetate,they were separated on a Allure PFP Propyl（100 mm×2.1 mm,5 μm） with a mobile phase of methanol-20 mmol/L ammonium acetate using gradient elution.For mass spectrometric detection,electrospray ionization（ESI＋）in positive mode was elected and the data was collected using multiple-reaction monitoring（MRM）.Results The linearity was good（r〉0.995 0） and the limit of detection was 2 ng/m L or 2 ng/g for triptolide and wilforlide A.The recovery was 61.08%-102.98%.The intra-day and inter-day precision was less than 12.58% for each biological sample,and the accuracy was 90.61%-105.80%.Conclusion This method is simple,convenient and good selective,and could be applied to analysis of triptolide and wilforlide A in different biological samples.And the method may provide technical support for forensic medicine identification,clinical diagnosis and treatment of tripterygium wilfordii Hook.f.poisoning.